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Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
Published on: March 13, 2015
Hepatitis C Core-Antigen Testing from Dried Blood Spots
Mia J Biondi1,2,3, Marjolein van Tilborg4,5, David Smookler6,7
1Toronto Centre for Liver Disease, Toronto General Hospital, University Health Network, University of Toronto, Toronto, ON M5G 2C4, Canada. mia.biondi@mail.mcgill.ca.
Insights
Dried blood spot (DBS) testing for hepatitis C virus (HCV) antibodies and core antigen shows high accuracy across various temperatures. This method offers a viable alternative to HCV RNA testing, improving accessibility and reducing costs for HCV screening.
Area of Science:
- Virology
- Immunology
- Diagnostic Medicine
Background:
- Hepatitis C virus (HCV) screening requires accessible and cost-effective diagnostic methods.
- Dried blood spot (DBS) collection offers a promising alternative for sample collection and transportation.
Purpose of the Study:
- To evaluate the diagnostic accuracy of HCV antibody (HCV-Ab) and HCV core antigen (core-Ag) testing using DBS cards under various temperature conditions.
- To compare the performance of DBS testing with venous and capillary blood samples.
Main Methods:
- Blood samples from 68 chronic HCV patients were collected on DBS cards and stored for one week at temperatures ranging from -80 °C to 37 °C.
- Eluted samples were tested for HCV-Ab and core-Ag.
- A head-to-head comparison of venous vs. capillary samples and single vs. double DBS spots was performed.
Main Results:
- High sensitivity for HCV-Ab detection was observed across all tested temperatures.
- Core-Ag sensitivity remained high (91-94%) across different storage temperatures.
- No significant difference in core-Ag sensitivity was found between venous and capillary samples, with improved sensitivity using two DBS spots.
Conclusions:
- HCV-Ab and core-Ag testing using DBS cards demonstrates high diagnostic accuracy and stability under various temperature conditions.
- DBS testing is a reliable alternative to HCV RNA testing, particularly in settings where accessibility and cost are considerations.
- Further validation supports the use of DBS cards for expanded HCV screening programs.
Abstract:
In order to expand hepatitis C virus (HCV) screening, a change in the diagnostic paradigm is warranted to improve accessibility and decrease costs, such as utilizing dried blood spot (DBS) collection. In our study, blood from 68 patients with chronic HCV infection was spotted onto DBS cards and stored at the following temperatures for one week: -80 °C, 4 °C, 21 °C, 37 °C, and alternating 37 °C and 4 °C; to assess whether temperature change during transportation would affect sensitivity. Sample was eluted from the DBS cards and tested for HCV antibodies (HCV-Ab) and HCV core antigen (core-Ag). HCV-Abs were detected from 68/68 DBS samples at -80 °C, 4 °C, 21 °C, and 67/68 at 37 °C and alternating 37 °C and 4 °C. Sensitivity of core-Ag was as follows: 94% (-80 °C), 94% (4 °C), 91% (21 °C), 93% (37 °C), and 93% (37 °C/4 °C). Not only did temperature not greatly affect sensitivity, but sensitivities are higher than previously reported, and support the use of this assay as an alternative to HCV RNA. We then completed a head-to-head comparison (n = 49) of venous versus capillary samples, and one versus two DBS. No difference in core-Ag sensitivity was observed by sample type, but there was an improvement when using two spots. We conclude that HCV-Abs and core-Ag testing from DBS cards has high diagnostic accuracy and could be considered as an alternative to HCV RNA in certain settings.
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